Small Molecule Enhancers of Paneth Cell Function and Differentiation

Abstract

Leucine-rich repeat-containing G-protein coupled receptor 5-positive (LGR5+) intestinal cells are contacted with an inhibitor of exportin 1 (XPO1), thereby producing functionally differentiated intestinal cells. The LGR5+ cells can also be contacted with a Wnt agonist. The LGR5+ cells can also be contacted with a Notch inhibitor.

1 . A method of differentiating leucine-rich repeat-containing G-protein coupled receptor 5-positive (LGR5 + ) intestinal cells, the method comprising contacting LGR5 +  cells with an inhibitor of exportin 1 (XPO1), thereby producing functionally differentiated intestinal cells. 
     
     
         2 . The method of  claim 1 , wherein the inhibitor of XPO1 is KPT-330. 
     
     
         3 . The method of  claim 1 , wherein the inhibitor of XPO1 is KPT-8602. 
     
     
         4 . The method of  claim 1 , wherein the inhibitor of XPO1 is Leptomycin B. 
     
     
         5 . The method of  claim 1 , further comprising contacting the LGR5 +  cells with a Wnt agonist. 
     
     
         6 . The method of  claim 5 , wherein the Wnt agonist is CHIR99021. 
     
     
         7 . The method of  claim 1 , further comprising contacting the LGR5+ cells with a Notch inhibitor. 
     
     
         8 . The method of  claim 7 , wherein the Notch inhibitor is DAPT. 
     
     
         9 . The method of  claim 1 , wherein the functionally differentiated intestinal cells are Paneth cells. 
     
     
         10 . The method of  claim 1 , wherein the functionally differentiated intestinal cells are CD24-mid/LYZ +  cells. 
     
     
         11 . The method of  claim 10 , wherein the CD24-mid/LYZ +  cells are Paneth cells. 
     
     
         12 . The method of  claim 1 , wherein the LGR5 +  intestinal cells are intestinal stem cells. 
     
     
         13 . The method of  claim 1 , wherein the functionally differentiated intestinal cells secrete greater quantities of lysozyme compared to the LGR5 +  intestinal cells. 
     
     
         14 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express any combination of one or more of human lysozyme (LYZ), a human alpha defensin (DEFA), human matrix metalloproteinase-7 (MMP-7), and cluster of differentiation 24 (CD24). 
     
     
         15 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express human lysozyme (LYZ). 
     
     
         16 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express a human alpha defensin (DEFA). 
     
     
         17 . The method of  claim 16 , wherein the human alpha defensin is human alpha defensin 5 (DEFA5) or human alpha defensin 6 (DEFA6). 
     
     
         18 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express mRNA for any combination of one or more of human lysozyme (LYZ), a human alpha defensin (DEFA), human matrix metalloproteinase-7 (MMP-7), and cluster of differentiation 24 (CD24). 
     
     
         19 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express mRNA for human lysozyme (LYZ). 
     
     
         20 . The method of  claim 1 , wherein the functionally differentiated intestinal cells express mRNA for a human alpha defensin (DEFA). 
     
     
         21 . The method of  claim 20 , wherein the mRNA for human alpha defensin is mRNA for human alpha defensin 5 (DEFA5) or mRNA for human alpha defensin 6 (DEFA6). 
     
     
         22 - 26 . (canceled) 
     
     
         27 . A method of treating graft-versus-host disease, inflammatory bowel disease, Crohn's disease, necrotizing enterocolitis, or intestinal inflammation in an individual in need thereof, the method comprising administering an effective amount of an inhibitor of exportin 1 (XPO1) to intestinal cells of the individual. 
     
     
         28 - 36 . (canceled)